The localization of TLR4 was determined by immunocytochemistry. Paraffin histologic sections were dewaxed with xylene and then rehydrated in graded concentrations of ethanol. Sections were incubated with rabbit anti-mouse TLR4 antibodies (eBioscience, San Diego, CA) overnight at room temperature. Secondary swine anti-rabbit antibodies (DAKO, Copenhagen, Denmark) conjugated to streptavidin-biotin horseradish peroxidase were added for 30 minutes. Staining was revealed using a HRP-Dab Staining Kit (Vector Laboratories, Burlingame, CA). Densitometric analysis was performed using MetaMorph/Evolution MP 5.0/BX51 Image analytical system (Beijing, China).
Hrp dab staining kit
Manufactured by Vector Laboratories
Sourced in Canada, United States
The HRP-DAB Staining Kit is a laboratory product used for the visualization of specific target proteins or molecules in biological samples. The kit utilizes horseradish peroxidase (HRP) enzyme and 3,3'-diaminobenzidine (DAB) as a chromogenic substrate to produce a brown-colored reaction product, indicating the presence and localization of the target of interest.
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Lab products found in correlation
2 protocols using hrp dab staining kit
1
Immunohistochemical Localization of TLR4
2
Histological and Immunohistochemical Analysis
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Samples of the distal colon, liver, and epididymal fat from each animal underwent a histological examination and immunohistochemical examination. These organs were fixed in 10% (v/v) neutral-buffered formalin, dehydrated in ethanol, and embedded in paraffin. The sections were sliced to a 5-μm thickness for each sample. Hematoxylin and eosin (H&E) staining and immunohistochemistry were performed using the standard protocol and used CD56 antibody (M7304, DAKO, Carpinteria, CA, USA), HRP-conjugated monoclonal secondary goat anti-mouse IgG (K4006, DAKO) and a horseradish peroxidase-diaminobenzidine (HRP-DAB) staining kit (SK-4100, VECTOR Lab., Burlingame, CA, USA). More detail information is provided in the supplement methods.
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